Analysis of starch swelling power in Australian breeding lines of hexaploid wheat (Triticum aestivum L.)

Starch is the major component of wheat (Triticum aestivum L.) grain and is composed of two large glucan molecules, amylose and amylopectin. The ratio between the two polymers types influences the water absorbing properties of starch upon heating, and thus affects the end-use of grain and purified starch. In this study, we evaluated the starch swelling power (SSP) values in seven wheat populations developed from crosses involving low-SSP lines. Analysis of starch produced by the F₂ generation plants showed that the largest SSP variation (11.4–16.2) and lowest SSP mean (13.9) was obtained for a population derived from doubled haploid lines SM1028 (SSP = 14.5) and VK306 (SSP = 13.6). The population of 360 lines was advanced by single seed descent to the following generations for further studies. Starch analysis of grain produced by F₄ generation lines in two field locations during 2006 and in a greenhouse environment during 2005 showed that SSP values were relatively stably inherited. The average broad-sense heritability was 73% and significant (P < 0.001) genotype × genotype and genotype × environment interactions were seen. Starches with the highest and lowest SSP values were inversely related to amylose concentration determined by high pressure liquid chromatography (HPLC)–size exclusion chromatography (SEC) of debranched starch. Developed lines with the lowest SSP values surpassed 40% in apparent amylose concentration. The study illustrates that screening for SSP in early generations can be used to develop wheat lines with desired starch swelling characteristics.     

The Null-4A Allele at the Waxy Locus in Durum Wheat affects Pasta Cooking Quality

Granule-bound starch synthase, also known as the waxy protein catalyses the synthesis of amylose in wheat endosperm starch. In durum wheats, the genes encoding GBSS are present at the two Wx loci on chromosome 7A and 4A (a segment of 7B that has been translocated). Several null Wx-B1 (missing GBSS protein from chromosome 4A) durum lines were produced from crosses with null-4A bread wheats backcrossed to durum wheats. Semolina milled from 4 normal and 7 null-4A durum wheat lines grown over two seasons (1999 and 2000) in South Australia were analysed for amylose content, starch pasting properties as measured by the Rapid Viscoanalyzer (RVA), swelling power and starch damage, protein content and electrophoretic protein analysis. Spaghetti was prepared with a micro-scale extruder and the cooked pasta evaluated for cooking loss, firmness, stickiness and water absorption. The null-4A lines had significantly lower (ca. 5%) amylose content, higher starch peak viscosities and semolina swelling power. The pasta derived from the null-4A lines had lower cooking loss and in 1999 was more adhesive than the non-waxy lines. Cooking loss was correlated with amylose content, peak starch viscosity, swelling power of semolina and cooked pasta adhesiveness. Semolina swelling power was highly correlated with RVA peak viscosity. Waxy durum wheats appear to have an advantage over the normal types in terms of lower cooking loss, widely used as an indicator of pasta cooking quality.     

Survey of amylose content in Secale cereale, triticum monococcum, T. turgidum and T. tauschii

A standard method for determining apparent amylose content was modified for use with samples of 2–3 cereal seeds. Starch was extracted by soaking the seeds overnight in dilute ammonia solution, grinding in NaCl solution in a microfuge tube with an appropriate pestle and decanting the starch slurry. The starch was then washed successively in acetic acid, ethanol and acetone. The amylose content was determined by dispersing 5·0 mg of the starch in ethanol, adding NaOH solution, heating until a clear gel was formed, diluting, neutralising with citric acid, staining with iodine and reading in a spectrophotometer at 620 nm. The method provided a very strong correlation with a standard method for much larger samples. The apparent amylose content was determined for 1083 accessions of Triticum monococcum (einkorn), T. turgidum (emmer), T. tauschii and Secale cereale (rye). The 10 extreme accessions of each species were re-analysed a further three times and the two most extreme individual lines were selected for genetic studies. Apparent amylose content of T. monococcum ranged from 15 to 28%; that of T. turgidum, 19–31%; T. tauschii, 21–34%; and rye 12–28%. These ranges are considered sufficiently broad to allow amylose content to be further diversified through breeding.     

Effects of two novel Wx-A1 alleles of common wheat (Triticum aestivum L.) on amylose and starch properties

Common wheat (Triticum aestivum L.) has three Wx proteins (Wx-A1, Wx-B1, and Wx-D1), which are granule-bound starch synthases I and are responsible for the amylose synthesis of flour starch. The effects of two novel Wx-A1 proteins identified by gel electrophoresis on amylose content and starch properties were analyzed. The variant Wx-A1 protein coded by the Wx-A1i allele was present in smaller amounts and produced less amylose (7.3%) compared to standard lines with the Wx-A1a allele (21.5%). Wx-A1i generated altered starch properties; greater swelling power (SP), glucoamylase digestibility, starch paste clarity, and gelatinization enthalpy in differential scanning calorimetry. The starch from Wx-A1i also showed an altered pasting profile on a Rapid Visco-Analyzer, greater peak viscosity, smaller final viscosity, and lower pasting temperature. The Wx-A1i allele is a novel genetic resource for reducing amylose content in wheat. The other Wx-A1 protein coded by the Wx-A1j allele showed a more basic isoelectric point compared to Wx-A1a on an electrophoretic gel. The amylose content of Wx-A1j did not differ from standard Wx-A1a. Starch SP, paste clarity, and glucoamylase digestibility also suggested that Wx-A1j produced amylose as much as Wx-A1a.     

Water stress during grain development affects starch synthesis,composition and physicochemical properties in triticale

Triticale, a man-made cereal crop developed from a cross between wheat and rye, has excellent agronomic traits to produce starch for bioindustrial applications. The effects of different levels of water stress on expression of starch synthesis genes and starch composition and physicochemical properties were investigated in this study. Three triticale varieties from 5 days post-anthesis were treated with three levels of water stress: low water stress (LWS) at 55–60% of soil moisture, moderate water stress (MWS) at 30–35% soil moisture and severe water stress (SWS) at 10–15% soil moisture. Water stress led to a significant reduction in seed weight at SWS (35–45%). A decrease in starch content was noticeable from MWS onwards and the values were decreased by 42–55% at SWS across all varieties. Such decrease was associated with the reduced expression of starch synthesis genes at 19 days of water stress (DWS). MWS favoured an increase of amylose proportion in triticale starch and it was accompanied by a significant up-regulation of GBSSI expression throughout the grain development. Triticale starch synthesized under water stress showed a reduced population of small granules and an increase of A-type to B-type ratio. SWS caused pitting on starch granules but did not alter the biconcave disc shape of mature granules. An inverse relationship between water stress and a range of starch gelatinization temperature was observed and the MWS environment specifically decreased the peak temperature (Tp) and increased the enthalpy. Our results signify that starch morphology, composition and physicochemical properties in triticale grains could be altered if triticale is grown under drought conditions.     

Ae显性突变体对玉米农艺性状及直链淀粉含量的影响分析

玉米直链淀粉是重要的食品和工业原料,高直链淀粉性状的产生受多种酶和基因-环境等因素的共同影响。玉米ae基因（amylose extender）突变导致子粒中直链淀粉含量增高、子粒表面皱缩、淀粉颗粒呈不规则形状、尺寸不均匀且表面基质蛋白增多,用于玉米育种后获得的品种直链淀粉含量升高,产量显著降低。通过将显性Ae突变体与16份优质玉米自交系杂交,对F₁代植株进行开花期、株型、产量性状调查,分析F₁代子粒总淀粉、直链淀粉的含量。结果表明,Ae突变体与不同的自交系杂交,F₁代直链淀粉含量及产量的变化存在明显差异,与基因型有密切关系,以Ae突变体和W16F41、CA240为亲本杂交,可显著提高直链淀粉含量同时产量变化最小。根据F₁代表现筛选直链淀粉含量提高、产量变化较小的双亲配制杂交种,可加速高直链淀粉玉米新品种培育进程。     

Water stress during grain development affects starch synthesis,composition and physicochemical properties in triticale

Triticale, a man-made cereal crop developed from a cross between wheat and rye, has excellent agronomic traits to produce starch for bioindustrial applications. The effects of different levels of water stress on expression of starch synthesis genes and starch composition and physicochemical properties were investigated in this study. Three triticale varieties from 5 days post-anthesis were treated with three levels of water stress: low water stress (LWS) at 55–60% of soil moisture, moderate water stress (MWS) at 30–35% soil moisture and severe water stress (SWS) at 10–15% soil moisture. Water stress led to a significant reduction in seed weight at SWS (35–45%). A decrease in starch content was noticeable from MWS onwards and the values were decreased by 42–55% at SWS across all varieties. Such decrease was associated with the reduced expression of starch synthesis genes at 19 days of water stress (DWS). MWS favoured an increase of amylose proportion in triticale starch and it was accompanied by a significant up-regulation of GBSSI expression throughout the grain development. Triticale starch synthesized under water stress showed a reduced population of small granules and an increase of A-type to B-type ratio. SWS caused pitting on starch granules but did not alter the biconcave disc shape of mature granules. An inverse relationship between water stress and a range of starch gelatinization temperature was observed and the MWS environment specifically decreased the peak temperature (Tp) and increased the enthalpy. Our results signify that starch morphology, composition and physicochemical properties in triticale grains could be altered if triticale is grown under drought conditions.     

Effect of jet-cooked wheat gluten/lecithin blends on maize and rice starch retrogradation

Vital wheat gluten and lecithin (GL) (50:50, w/w) were dry blended in a coffee grinder and a 9.5% (w/v) aqueous slurry was jet-cooked (steam pressures of 65 psi/g inlet and 40 psi/g outlet) to disaggregate wheat gluten and facilitate better dispersion of the two components. The jet-cooked material was freeze-dried and stored at 0 °C for future use. The GL blend was added to pure food grade common maize and rice starch at concentrations of 0 (control), 6, 11, 16, and 21%. Starch gelatinization and retrogradation temperature transitions were determined using Differential Scanning Calorimetry (DSC). From the DSC profiles, the change in the ΔH value was used as an indication of starch retrogradation, where a higher ΔH value indicated higher retrogradation. The ΔH values of the blends at 4 °C had higher values than the −20 °C and the ambient (25 °C) storage temperatures. Overall, the 21% GL/starch blends reduced retrogradation by 50%. The lower amylose content of rice starch relative to maize starch was reflected in Rapid Visco Amylograph (RVA) measurements of peak viscosity, and similarly, Texture Analyzer (TA) measurements indicated that maize starch gel is firmer than rice starch gel. Retrogradation was also evaluated by observing G′, the shear storage modulus, as a function of time after running a standard pasting curve. Using this method, it appears that GL has a significant effect on maize starch retrogradation, since low concentrations (<0.4%, w/w) reduced G′ up to 40%. The opposite behavior was seen in rice starch, where G′ increased directly with added GL. It appears that the amylose level in the rice starch is too low to be affected by the GL, and the increase seen in G′ is most likely due to added solids.     

蔗糖合成酶(SUS)基因的SNP标记开发及其与淀粉性状的关联分析

通过直接测序法获得玉米亲本(H21和紫糯96-619)sus1基因全长序列并进行亲本间序列比对,得到120个SNP位点,通过高分辨率熔解曲线(HRM)对300个近等基因系H21 BC5F2:3进行SNP位点分型,并将后代的基因型与群体的直链淀粉含量和淀粉糊化特性进行关联分析。结果表明,marker 31与直链淀粉含量和淀粉崩解值、峰值时间、糊化特性均连锁紧密;marker 17与直链淀粉含量和淀粉的峰值时间、淀粉最终黏度均连锁紧密;marker12与直链淀粉含量、淀粉的峰值时间连锁紧密。     

Amylolysis of wheat starches. I. Digestion kinetics of starches with varying functional properties

The susceptibility of wheat (Triticum aestivum L.) starches to hydrolysis by pancreatic α-amylase in vitro was investigated using a series of 35 starches with slightly enriched amylose content within a narrow range (36–43%), but widely differing functional properties. After 2 h of incubation with α-amylase, native starch granules were digested to different extents, but there were no differences between any of the starches once they were gelatinized. Cooling the starch for 72 h at 4 °C after cooking reduced the susceptibility of all of the starches to enzymic digestion by a similar extent, whereas addition of monopalmitin decreased the digestibility of the starches that contained amylose, but did not affect the digestibility of waxy starches that were also included in the study. Amylopectin chain length distribution of partly digested starch granules displayed increased proportion of short and medium chains and decreased proportion of long chains in comparison to native granules. Separated large (A) and small (B) starch granules from three of the starches differed significantly in their susceptibility to in-vitro digestion. A predictive model of the susceptibility of starch in the different forms was developed from the physico-chemical and functional properties of the starches.     

Effect of wx genes on amylose content,physicochemical properties of wheat starch,and the suitability of waxy genotype for producing Chinese crisp sticks

The effect of wx genes on amylose content, physicochemical properties of wheat starches, and the quality of Chinese crisp stick were investigated using near-isogenic lines (NILs) with null wx alleles in Yangmai 17 and Yangmai 01-2 backgrounds. wx genes showed significant effects on amylose content and other traits. The triple-null genotype had the lowest amylose content among eight genotypes, followed by double-null, single-null, and wild-type genotypes. The triple-null also showed lower flour yield, higher percentage of type B-granules on a volume basis and higher crystallinity than non-waxy genotypes, and showed significant differences in all pasting and thermal transition parameters compared to non-waxy genotypes, except for degree of retrogradation at day 14. For the quality of Chinese crisp stick, the hardness, crispness, fracturability, and specific volume of waxy genotype were 3.91 kg, 11.0, 1.85 mm and 104.4 ml, whereas the corresponding ranges for non-waxy genotypes were 5.39–5.70 kg, 0.5–0.9, 0.69–0.86 mm and 49.5–57.6 ml, respectively, in Yangmai 17 background. This indicates that waxy genotypes showed significantly better crisp stick quality than non-waxy genotypes. A similar trend was also observed in Yangmai 01-2 background. This indicated the potential utilization of waxy wheat for producing traditional products.     

糯性普通小麦的产生及其淀粉特性研究

以江苏白火麦（缺失Wx-Dl)和关东107（缺失Wx-Al和Wx-Bl）为亲本配制杂交组合，采用花粉和籽粒剖面染色、蛋白质电泳进行筛选和鉴定，人工创造得到自然界不存在的糯性小麦，并对其直链淀粉含量及淀粉品质性状进行研究。结果表明，所得到的两个糯性小麦吕系直链淀粉含量很低（小于1％），其糊化特性、膨胀势特性亦与普通小麦品种不同；RVA高峰粘度明显低于其亲本，其高峰粘度出现较快，而且反弹值不明显；其淀粉膨胀能力远大于亲本。     

高直链淀粉小麦西农836淀粉理化性质及消化特性分析

淀粉是人体摄入碳水化合物的主要来源,经消化后以葡萄糖的形式被人体吸收,其消化特性与人体健康密切相关。为探究小麦品种西农836淀粉理化性质与消化特性的关系,以商品粉金沙河、香雪、金龙鱼作为对照,测定其与西农836中直链淀粉含量、淀粉粒度和结晶度、糊化特性以及体外消化率的差异,并在小鼠体内模拟消化。结果表明,与金龙鱼、金沙河、香雪相比,西农836淀粉中直链淀粉含量和B型淀粉粒含量最高,与前3者差异显著（P<0.05）;西农836淀粉的峰值温度最高,峰值黏度和最终黏度最低;淀粉体外消化试验结果显示,西农836的快消化淀粉（RDS）、总消化淀粉（RDS+SDS）含量最低,抗性淀粉（RS）含量最高。小鼠体内消化试验结果表明,与金沙河相比,灌食西农836淀粉的小鼠血糖峰值较低,说明高直链淀粉小麦西农836有利于延缓消化和控制血糖升高。相关性分析结果显示,直链淀粉含量与抗性淀粉含量呈显著正相关（P<0.05）。     

Physicochemical Studies on Resistant StarchIn VitroandIn Vivo

Resistant starch (RS), producedin vitroby hydrolysis of retrograded pea starch gels and amylose gels by porcine pancreaticalpha-amylase, was characterised by X-ray diffraction, size exclusion chromatography and methylation analysis. These techniques showed that RSin vitroconsisted of semi-crystalline, mostly linear material that was present in two main molecular size subfractions (DP_n>100 andDP_n20–30) with a third, minor subfraction (DP_n≤5). The extent of retrogradation of amylose was found to be of primary importance in determining the RS content of starch. Analysis ofin vivoRS, recovered during an ileostomy study, produced results that were similar to those obtained from RSin vitro. Anin vitromodel for the structure of resistant starch is proposed.     

Formulating low glycaemic index rice flour to be used as a functional ingredient

Amylose and resistant starch (RS) content in rice flour were manipulated. The experiment was conducted using a full factorial design. Rice flour with average amylose content of 20 and RS content of 0.5 g/100 g dry sample was fortified with pure amylose from potato and high RS modified starch to reach the final amylose content of 30, 40 and 50 and RS content of 2, 4 and 6 g/100 g dry sample. The fortified rice flours were examined for their gelatinisation properties, in-vitro enzymatic starch digestion and gel textural properties. It was found that amylose and RS significantly affect all the fortified rice flour properties (p < 0.05). High amylose and RS improved starch digestion properties, reducing the rate of starch digestion and lowering the glycaemic index (GI) values. Amylose had a more pronounced effect on the fortified rice starch properties than RS. In this study, the fortified rice flour which contained amylose and RS of approximately 74 and 9 g/100 g dry sample respectively was used to produce rice noodles. The noodles exhibited low GI values (GI < 55). However, amylose and RS affected the textures of rice noodles providing low tensile strength and break distance (extensibility).     

Discovery of lunasin peptide in triticale (X Triticosecale Wittmack)

Lunasin is a novel, cancer-preventive, anti-inflammatory and cholesterol-reducing peptide that was originally isolated from soy and later from barley, wheat and rye. We report the first discovery of lunasin in triticale (X Triticosecale Wittmack). Moreover, we report first data of lunasin content in winter rye and wheat genotypes grown in Northern Europe. These data are novel as previously published data on finding of lunasin in cereals were obtained in genotypes grown in Korea. Lunasin content was uncovered using a previously published procedure for isolation from cereals and identified by LC-MS/MS assay. We found that triticale was the most lunasin-rich cereal, with the tested genotypes displaying the following trend in lunasin content: genotype 0002-26 > Dinaro > DSGU 10/94 > 0213-22 > 0317-14 > 0006-31. The greatest lunasin content was 6.46 mg/g in the grain of triticale genotype 0002-26. In comparison, the highest lunasin content in rye variety Dankovske Diament was 1.5 mg/g, and the highest lunasin content in the winter wheat variety Fredis was 0.23 mg/g. We conclude that triticale can play a significant role as functional food, with great potential for the use of triticale products in human and animal diets.     

In VitroDigestion of Wheat Microstructure with Xylanase and Cellulase fromTrichoderma reesei

Resistant starch (RS), producedin vitroby hydrolysis of retrograded pea starch gels and amylose gels by porcine pancreaticalpha-amylase, was characterised by X-ray diffraction, size exclusion chromatography and methylation analysis. These techniques showed that RSin vitroconsisted of semi-crystalline, mostly linear material that was present in two main molecular size subfractions (DP_n>100 andDP_n20–30) with a third, minor subfraction (DP_n≤5). The extent of retrogradation of amylose was found to be of primary importance in determining the RS content of starch. Analysis ofin vivoRS, recovered during an ileostomy study, produced results that were similar to those obtained from RSin vitro. Anin vitromodel for the structure of resistant starch is proposed.     

半粒小麦胚乳直链淀粉含量测定方法的研究

小麦直链淀粉含量对面条品质有较大的影响，并已被育种家们作为优质面条小麦选育的品质指标之一．测定半粒小麦直链淀粉的含量对小麦品质育种早代材料的筛选尤为重要。本实验对半粒小麦样品的前处理及其直链淀粉含量的碘比色测定法进行了研究，提出了半粒小麦样品的直链淀粉含量的分析方法，并将用该法测定的半粒小麦样品的直链淀粉含量与常规的碘比色法测得的群体小麦样品的直链淀粉含量进行了简单线性相关分析，结果表明，两种方法的分析结果呈极显著相关(r=O．9508)。本研究所得的半粒小麦样品的直链淀粉含量测定方法是一种准确、简便和有效的测定方法。     

Starch Properties of a Bread Wheat (Triticum aestivum L.) Mutant with an Altered Flour-pasting Profile

Ethyl methanesulphonate treatment of seeds from cultivar Kanto 107 lacking Wx-A1 and Wx-B1 proteins induced a mutant, named K107Afpp4, showing an altered flour-pasting profile, i.ea markedly increased viscosity at a low temperature and a decreased peak time in Rapid Visco Analyser measurement. The colour of the iodine complex of the isolated starch from the mutant remained bluish-purple but its λ_maxdecreased to 584 nm from 601 nm shown by the wild type Kanto 107. The absorbance at 680 nm (blue value) of the mutant starch decreased to 0·206 from 0·324 for Kanto 107. The amylose content of K107Afpp4 starch was 12·7% measured by concanavalin A method, compared to 21·9% for the wild type. The apparent amylose content of the mutant starch measured by amperometric titration was 15·9% while that of Kanto 107 was 25·3%. Gel permeation chromatography showed that the mutant starch has a decreased amount of lower molecular weight fractions than starch from the wild type. Differential scanning calorimetry indicated that mutant starch has a higher gelatinisation peak temperature, 61·8 °C, than the wild type, 59·5 °C. The mutant will be useful as a genetic resource for altering wheat starch/flour properties and for clarifying the relationship between amylose content and wheat quality.